Journal: iScience

Article Title: CFP1 promotes B cell class switch recombination through the regulation of S region germline transcription and proliferation

doi: 10.1016/j.isci.2026.116255

Figure Lengend Snippet: CFP1 is critical for class-switch recombination in activated B cells (A) Representative flow cytometry plots and ratio of IgA CSR efficiency in CFP1-deficient CH12F3 cells under CIT stimulation for 3 days ( n = 3). (B and C) Representative flow cytometry plots and ratio of IgG1 and IgA switching in Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells under LPS/IL-4/TGF-β/IL-5/δ-glucoside ( n = 4) (B) or LPS/IL-4 conditions (n = 6–7) (C). (D) Representative flow cytometry plots and ratio of the IgG3 switching in Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells under LPS conditions (n = 7–8). (E) Schematic of DC-PCR with genomic DNA from stimulated B cells. (F) Detection of deletional Sμ-Sγ1 joining by DC-PCR in stimulated splenic B cells from Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre ( n = 3), Chrnb1 served as the internal control. For all panels showing quantitative data, data are shown as mean ± SEM. p values by unpaired t test (two-tailed).

Article Snippet: Cd21 cre , The Jackson Laboratory , Cat# 006368.

Techniques: Flow Cytometry, Control, Two Tailed Test

Journal: iScience

Article Title: CFP1 promotes B cell class switch recombination through the regulation of S region germline transcription and proliferation

doi: 10.1016/j.isci.2026.116255

Figure Lengend Snippet: CFP1 is required for optimal S region GLT and AID targeting (A and B) Illustration shows the location of RT-qPCR primers for spliced and initiating Igh switch transcripts (A) and quantification of these transcripts (B) in Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells ( n = 3). (C) RT-qPCR analysis of Aicda gene in Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells stimulated under LPS/IL-4 for 2 days ( n = 6). (D) Schematic of the 5′ Sμ SHM-seq workflow in splenic B cells from Aicda −/− , Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre mice (Upper). Mutation profiles in the above B cells stimulated with LPS/IL4 for 4 days (Lower). Green bars indicate SEM. Positions of AGCT and WRC (W: A/T; R: A/G) motifs are marked with orange and yellow bars, respectively. (E) Normalized mutation frequency in Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre mice ( n = 3). (F) Gene expression in splenic B cells stimulated with LPS/IL-4 for 2 days by mRNA-seq ( n = 3). For all panels showing quantitative data, data are shown as mean ± SEM. p values by unpaired t test (two-tailed).

Article Snippet: Cd21 cre , The Jackson Laboratory , Cat# 006368.

Techniques: Quantitative RT-PCR, Mutagenesis, Gene Expression, Two Tailed Test

Journal: iScience

Article Title: CFP1 promotes B cell class switch recombination through the regulation of S region germline transcription and proliferation

doi: 10.1016/j.isci.2026.116255

Figure Lengend Snippet: CFP1 knockout does not affect DSB end joining (A) DSB repair-related gene expressions in splenic B cells stimulated with LPS/IL-4 for 2 days assessed by mRNA-seq ( n = 3). (B) Schematic of Cas9/sgRNA-mediated IgG1 switching analysis. (C and D) Representative flow cytometry plots (C) and quantification (D) of the IgG1 class switching in CH12F3 cell nucleofection of sgRNAs targeting Sμ and Sγ1 for 48 h ( n = 7). (E, F, and G) Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells stimulated with LPS/IL-4 for 4 days ( n = 3). (E) Distribution of Sμ-Sγ1 and Sμ-Sε junctions recovered from stimulated B cells. Numbers in parentheses denote total unique junctions in the indicated regions. The area between the dotted lines indicates the core S region. (F) The ratio of inversional versus deletional junctions in Sγ1 and Sε regions (left) and percentage of end resection in Sγ1 and Sε regions (right) (G). The percentage of MH utilization within Sγ1 and Sε regions. For all panels showing quantitative data, data are shown as mean ± SEM. p values by unpaired t test (two-tailed) in (A) and (F). p values by Dunnett’s multiple comparisons test in (D).

Article Snippet: Cd21 cre , The Jackson Laboratory , Cat# 006368.

Techniques: Knock-Out, Flow Cytometry, Two Tailed Test

Journal: iScience

Article Title: CFP1 promotes B cell class switch recombination through the regulation of S region germline transcription and proliferation

doi: 10.1016/j.isci.2026.116255

Figure Lengend Snippet: Cfp1 ablation impairs cell proliferation (A) Representative flow cytometry plots and proliferation index analysis of CFSE-labeled Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells cultured with LPS/IL-4 for 96 h (n = 5–6). (B) Representative flow cytometry plots and the percentage of IgG1 class switching per cell division, tracked by CFSE dilution after 96 h of LPS/IL-4 stimulation (n = 5–6). (C) Representative flow cytometry plots and proliferation index analysis of CFSE-labeled Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells cultured with CD40/IL-4 for 96 h (n = 5–6). (D) Representative flow cytometry plots and the percentage of IgG1 per cell division, tracked by CFSE dilution after 96 h of CD40/IL-4 stimulation (n = 5–6). (E and G) Live cell counts of Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells at 0 h, 48 h, 72 h, and 96 h after LPS/IL-4 (E, n = 3) or CD40/IL4 stimulation (G, n = 3). (F and H) Cell-cycle analysis of splenic B cells stimulated with LPS/IL-4 (F, n = 4–5) or CD40/IL4 (H, n = 3) for 48 h. Bars represent the mean values in (B), (D), (F), and (H). For all panels showing quantitative data, data are shown as mean ± SEM. p values by unpaired t test (two-tailed).

Article Snippet: Cd21 cre , The Jackson Laboratory , Cat# 006368.

Techniques: Flow Cytometry, Labeling, Cell Culture, Cell Cycle Assay, Two Tailed Test

Journal: iScience

Article Title: CFP1 promotes B cell class switch recombination through the regulation of S region germline transcription and proliferation

doi: 10.1016/j.isci.2026.116255

Figure Lengend Snippet: Cfp1 regulates cell proliferation-related gene expression through H3K4me3 modification Cfp1 fl/fl and Cfp1 fl/fl Cd21 Cre splenic B cells are stimulated with LPS/IL-4 for 48 h for bulk mRNA-seq and CUT&Tag; n = 3. (A) Volcano plot of differentially expressed genes ( p value < 0.05, absolute log 2 FC > 0.5). (B) GSEA enrichment plot for pathways ranked by P .adjust. (C) Scatterplot showing the correlation between differential gene expression (DEGs in A and altered promoter H3K4me3 levels in Cfp1 -KO versus WT stimulated B cells. Pearson’s correlation coefficient is indicated. (D) GO enrichment in “both down” and “both up” genes shows the top five pathways ranked by q -values for each category. (E) Heatmap for mRNA and H3K4me3 signals of DEGs from B. (F) Genome browser tracks of mRNA and H3K4me3, for example, positive and negative cell proliferation-related genes. (G) Live cell counts of CH12F3 cells transduced with lentivirus expressing the Pcna or empty vector (EV) shRNA at 0 h, 24 h, 36 h, and 48 h cultured in vitro ( n = 3). (H) Representative flow cytometry plots and ratio of IgA CSR efficiency in EGFP + CH12F3 cells under CIT stimulation ( n = 3). Bars represent the mean ± SD in (G). For all panels showing quantitative data, data are shown as mean ± SEM. p values by Dunnett’s multiple comparisons test. p values based on 48 h data in (G). See also .

Article Snippet: Cd21 cre , The Jackson Laboratory , Cat# 006368.

Techniques: Gene Expression, Modification, Transduction, Expressing, Plasmid Preparation, shRNA, Cell Culture, In Vitro, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Association of CCL21 + CAFs with B-cell recruitment and TLS maturation in penile squamous cell carcinoma

doi: 10.1007/s00262-026-04348-5

Figure Lengend Snippet: Evaluation of TLS maturation and prognostic analysis of PSCC patients. A UMAP plot of the cell types identified in PSCC through spatial transcriptomic sequencing, depicting 7 distinct cell populations. B Dot plot displaying the expression of cell type-specific markers. The dot size represents the proportion of cells expressing the marker, while the colour intensity indicates the mean expression level. C Regions with AUCell scores for mTLS genes ( BCL6 , AICDA , CD38 , ICOS , CXCR5 , CXCL13 , and CD21 ) above the median were defined as mTLSs, whereas those below the median were classified as iTLSs. D Spatial visualization of AUCell scores for mTLS genes, where warmer colours indicate higher scores. E Spatial distribution of iTLSs (green) and mTLSs (orange) across different regions of PSCC tissues. F Representative images H&E staining and immunofluorescence staining for CD20 (green) and CD21 (red) in iTLSs and mTLSs. G GO enrichment analysis of upregulated genes in mTLS. Darker colours denote more significant enrichment of the pathway, and larger dots represent a greater number of genes involved. H Kaplan–Meier survival curves showing that patients with mTLSs experienced significantly longer overall survival than those with iTLSs, as determined using ssGSEA ( P < 0.05)

Article Snippet: The following antibodies were used: CD20 (Proteintech, 60,271–1-Ig, 1:500), CD21 (Proteintech, 28,206–1-AP, 1:250), ACTA2/α-SMA (Sigma, SAB5700835, 1:300), and CCL21 (Abcam, ab9851, 1:300).

Techniques: Sequencing, Expressing, Marker, Staining, Immunofluorescence